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This dissertation presents the synthesis and application of nanodiamond based materials for electrochemical biosensors. In this research work, nanodiamond particles have been used to prepare doped and undoped nanocrystalline diamond films, and conducting polymer composites for enhanced biosensing. The performance of the synthetized materials towards sensing applications was evaluated against glucose amperometric biosensing. Besides, cholesterol biosensing was attempted to prove the capabilities of the platform as a generic biosensing substrate. Biosensors have been proved to provide reliable detection and quantification of biological compounds. The detection of biological markers plays a key factor in the diagnosis of many diseases and, even more importantly, represents a major aspect in the survival rate for many patients. Among all of the biosensors types, electrochemical biosensors have demonstrated the best reliability to cost ratio. Amperometric biosensors, for example, have been used for decades as point of care sensing method to monitor different conditions such as glucose. Despite the amount the research presented, the sensitivity, selectivity, stability, low cost and robustness are always driving forces to develop new platforms for biosensor devices. In the first phase of this dissertation, we synthesized undoped and nitrogen doped nanocrystalline diamond films. The synthetic material was thoroughly studied using different material characterization techniques and taken through a chemical functionalization process. The functionalization process produced a hydrogen rich surface suitable for enzymatic attachment. Glucose oxidase was covalently attached to the functionalized surface to form the biosensing structure. The response of the biosensor was finally recorded following voltammetry and amperometric techniques under steady state and dynamic conditions. The experimental results demonstrated that conductivity induced by the doping process enhanced the sensitivity of the sensing structure with respect to the undoped substrate. Also, the functionalization procedure showed strong bonding to avoid enzyme leaching during the measurements. Later, in the second phase of this dissertation, the nanodiamond particles were used as filler for conducting polymer composites. The objective for developing these composite materials was to overcome the high resistivity observed for nanocrystalline films. The experimental results demonstrated that the inclusion of nanodiamond particles increased the sensitivity of the overall structure towards the quantification of glucose with respect to the nanocrystalline films and the bare polymer. Besides, the experiment showed a noticeable enhancement in the signal-to-noise ratio and the mechanical stability of the sensing platform due to the nanodiamond addition. The best structures from the previous experiments were further grafted with iron oxide nanoparticles to attempt signal amplification. Initial experiments with nanodiamond based composited showed similar current for low glucose concentrations for two different active electrochemical sensing areas. This observation indicates that more area is still available to transport signal and to enhance even further the sensing action. Oxidation of iron oxide nanoparticles after initial enzymatic decomposition of glucose has been proved to provide higher current for the same glucose concentration; thus, creating amplification effect for the signal. Finally, the toxicity of the nanomaterial synthesized during this dissertation was evaluated in mammalian cells. The advances in biosensing techniques indicate the potential application of amperometric platform for continuous implantable devices; hence, the toxicity of the materials becomes a key aspect of the platform design.

The ultimate goal of this dissertation research is to enhance the protein resistant nature of poly(ethylene oxide) (PEO) or poly(ethylene glycol) by introduction of a siloxane linker and to subsequently prepare coatings which prevent surface-induced thrombosis. The hydrophobicity and flexibility of the siloxane tether should impart both amphiphilicity and conformational mobility to the PEO chain to further decrease protein adhesion. Because adsorption of plasma (blood) proteins initiates the clotting process, coating surfaces based on these new PEO-silanes should prevent or significantly diminish thrombosis. Thus, these coatings would be extremely useful for bloodcontacting medical devices such as stents, grafts, arteriorintravenous shunts, and biosensors. Novel amphiphilic PEO-silanes were prepared with systematic variations to several key structural features, including: siloxane tether length, PEO segment length, and PEO architecture. Thus, PEO-silanes were prepared having the general formulas: a-(EtO)3Si(CH2)2-oligodimethylsiloxanen-block-[PEO8-OCH3] (n = 0, 4, and 13; linear architecture) and a-(EtO)3Si(CH2)2-oligodimethylsiloxanen-block-[PEOm-OCH3]2 (n = 0, 4, and 13; m = 6 and 12 branched architecture). The reactive triethoxysilane [(EtO)3Si-] group serves as the crosslinking or grafting moiety. The PEO segment is distanced from the (EtO)3Si- group by an oligodimethylsiloxane tether which is both hydrophobic and exhibits a high degree of chain flexibility. Crosslinked silicone coatings and surfacegrafted coatings were prepared with amphiphilic linear PEO-silanes (a-c). Crosslinked silicone coatings were also prepared with branched PEO-silanes (1a-3a and 1b-3b). All coatings showed improved resistance to common plasma proteins compared to silicone coatings. Furthermore, protein adsorption generally decreased with siloxane tether length. For crosslinked PEO-modified silicone coating systems based on linear (a-c) and branched PEO-silanes (1a-3a and 1b-3b), longer tethers enhanced PEO reorganization to the film-water interface to enhance protein resistance. In the absence of surface reorganization for surface grafted coatings prepared with linear PEO-silanes, longer siloxane tethers better inhibited protein adsorption despite a moderate decrease in graft density (sigma) and decrease in surface hydrophilicity. This indicates that longer siloxane tethers enhance the configurational mobility of the PEO segments to better repel proteins.

Lipid molecules are amphiphilic in nature consisting of a hydrophilic head group and hydrophobic hydrocarbon tails. The lipid bilayer consists of two layers of lipid molecules arranged with their hydrophobic tails facing each other and their hydrophilic head groups solvated by water. Lipid bilayers with hydrophilic polymer chains grafted onto the head groups have applications in various fields, such as stabilization of liposomes designed for targeted drug delivery, synthesis of supported bilayers for biomaterial applications, surface modification of implanted medical devices to prevent biological fouling and design of in vitro biosensors. The focus of this thesis lies in understanding the effects of polymer grafting on the thermodynamics and mechanical properties of lipid bilayers. Dissipative particle dynamics (DPD) has evolved as a promising method to study complex soft matter systems. The basic DPD algorithm, and its implementation are discussed in Chapter 2 of this thesis. It is important to achieve a tensionless state while studying phase transitions and deducing the mechanical properties of the bilayer. We proposed a modification of the Andersen barostat which can be incorporated in a DPD simulation to achieve the tensionless state as well as carry out simulations at a prescribed tension. In Chapter 3 of this thesis the effect of polymer grafting on single tailed lipid bilayers is studied. Simulations are carried out by varying the grafting fraction, Gf, defined as the ratio of the number of polymer molecules to the number of lipid molecules. At lowGf, the bilayer shows a sharp transition from the gel (Lβ) to the liquid crystalline (Lα) phase. This main melting transition temperature is lowered as Gf is increased. Corresponding to this, an increase in the area per head group is also observed. Above a critical value of Gf the interdigitated, LβI phase is observed prior to the main transition for the longer lipid tails. The analysis for two tailed lipids as a function of polymer chain length is extensively studied in Chapter 5. For the case of two tailed lipids, an intermediate interdigitated phase was not observed and the decrease in the melting temperature is more pronounced as the length of the polymer chain is increased. The scaling for fractional change in the area per head group, as well as the decrease in transition temperature as a function of polymer grafting are in good agreement with mean field theory predictions. The bending modulus (k) and area stretch modulus (kA) are essential for determining the shape and the mechanical stability of biological cells or lipid based vesicles. In simulations, the bending modulus k is evaluated from the Fourier transform of the out-of-plane fluctuations of the bilayer mid-plane. In Chapter 4 of this thesis, we illustrate that a surface representation based on Delanuay triangulation provides a robust parameter free representation of the bilayer surface. By evaluating the bending modulus for single tail lipids of different tail lengths, the continuum scaling relation d2 is verified. To our knowledge this is the first systematic investigation and verification of this scaling relationship using computer simulations. Using the continuum relation, =kAd2/ we find that α depends weakly on the tail lengths of the bilayer. Nevertheless we illustrate that a value of α=130 can be used to reliably estimate the bending modulus from the area stretch modulus for polymer free bilayers. Using our method, we are also able to capture the low q scalings and obtain the bending modulus of the gel (Lβ) phase. Grafted polymer was found to increase the value of the bending modulus for single tail lipids. Although the presence of polymer directly increases the area per head group, the suppressed height fluctuations dominate and the bending modulus increases for the single tail lipids. For two tail lipids a small decrease in the bending modulus was observed at low grafting fractions and short polymer chains. For large polymer lengths the bending modulus was found to increase monotonically.

After the advent of nanotechnology, self-assembly has become an active area of research, as it being one of the few efficient methods to generate ensembles of nanostructures. In this thesis, we present studies on two dimensional self-assembly of polymer grafted nanoparticle (PGNPs) and thin film modelling approach to understand the physics involved in the self-assembly mechanism of polymeric nanoparticles. The two dimensional, hierarchical assemblies of PGNPs are created from evaporating solution films spread at the air-water interface using Langmuir-Blodgett technique. A transition in the patterns is observed with increase in concentration which is followed by a remarkable re-entrance of initial patterns with further concentration increment. The pattern is long length scale network type at low and high concentrations whereas it is short length scale distribution of clusters at intermediate concentrations. Clusters are composed of lateral arrangement of individual PGNPs. The characteristics of clusters are tailored by changing various experimental conditions such as molecular weight of the grafted chains, concentration, temperature and evaporation rate. The patterns are unaffected by the transfer surface pressure, suggesting that the self-assembly occurs in the presence of solvent via solution thin film instabilities and the resulting structures of PGNPs are frozen upon complete evaporation. Films of neat polystyrene also exhibit similar morphology and transitions in pattern length scales with initial solution concentration as observed in PGNP films. This confirms that the self-assembly of PGNPs is driven by the intrinsic nature of the grafted polymer chains. Gradient dynamics model is employed to study the stability and dynamics of polymer solution thin films by incorporating Flory Huggins free energy and concentration dependent Hamaker constant. Dispersion curves obtained from linear stability analysis of thin film equations show existence of bimodal instability in the film that corresponds to dewetting and decomposition. Phase diagram spanned by concentration and Flory parameter indicate that the thin film instability transits from dewetting to decomposition and then re-enters to dewetting with increase in concentration of the solution. Using the material parameters of the PGNP thin films for linear stability analysis, experimental observations of bimodal length scale of patterns and re-entrant nature are well explained. Nonlinear simulations which are performed to capture the evolution of patterns in the film show that the decomposition progresses through different pathways depending upon the concentration of the solution. This is explained by analyzing the local variation of spinodal parameter (curvature of the free energy per unit area) in the film. The gradient dynamics model is extended to study the stability and dynamics of evaporating solution thin films. Nonlinear simulations demonstrate that the film undergoes evaporative thinning without any significant growth of dewetting or decomposition instability initially and becomes unstable at a certain intermediate thickness where the spinodal parameter of dewetting or decomposition changes the sign. The rupture of the film (dewetting) or the phase segregation (decomposition) occurs explosively and subsequently evaporation progresses till the film attains chemical equilibrium with the ambient vapour phase. Rate of evaporation significantly affect the intermediate thickness at which the patterns emerge and thereby determines the length scale of initial patterns and instability growth rate. Quasi-steady analysis and nonlinear simulations show that the length scales of patterns of dewetting and decomposition decrease with evaporation rate and exhibit a power law behaviour. Thin films in which the solvent quality drops down with confinement due to evaporation are modelled by assuming a simple functional dependence of Flory parameter on mean film thickness. Quasi-steady analysis demonstrates that the dominating instability of such films switches from dewetting to decomposition and then returns to dewetting with increase in the initial concentration of the solution. We note that even though the functional form of Flory parameter with confinement is not exact, it represents the essential nature of the expected variation. We presume that the phenomenon discussed above is quite generic and may manifest itself in many situations where thin films of colloidal solutions undergo a decrease in the solvent quality due to confinement effects resulting in a competition between spinodal dewetting and decomposition instabilities. This will result in a competition and interplay of the different instability scales and by choosing appropriate control parameters novel self-assembled patterns can be created.

This thesis presents the first reported grafted PPE - polycaprolactone-g-PPE; the first PPE based sensing model: biotinylated grafted PPE/streptavidin coated sphere; the first photocrosslinkable PPE ¨C allyloxy PPE; and the new mechanism which demonstrates morphology control on a single molecular level

Reported is a study of the crystallization behavior of syndiotactic polystyrene (sPS) and its copolymers (with 4-bromostyrene as the comonomer or with atactic polystyrene arms grafted on the comonomer sites) via three sets of experiments. The first involves the study of structural identification of negatively birefringent spherulites by means of polarized light microscopy (PLM) and scanning electron microscopy (SEM). Results indicated that the optically positive and optically negative spherulites have same morphological features. Differences in the optical texture are due entirely to differences in orientation of the (anisotropic) sheaf-like precursors: the rigid nature of crystalline lamellae renders incomplete development of spherical symmetry even at the axialitic size of tens of microns. In the second part, we propose a modified approach for more precise determination of the Tm* value by taking advantage of the dual-mode distribution of crystalline lamellae in analyzing small-angle X-ray scattering (SAXS) profiles. This method should be generally applicable to other semi-crystalline polymers with dual-mode distribution in lamellar thickness. Results from wide-angle X-ray diffraction (XRD) suggest the presence of ?'-to-?" phase transformation at ca. 264 oC; no indications for the previously proposed ?-to-? transformation are identified. We therefore conclude that the ?' form is truly metastable; the ?"-form is the entropically favored high temperature phase (with Tm* = 300 oC) whereas the more ordered ?' phase (with Tm* = 288 oC) is enthalpically favored at lower temperatures. In the third set of experiments, identification of effects of copolymerization has been studied via a combination of PLM, differential scanning calorimetry (DSC), XRD, SAXS, and transmission electron microscopy (TEM). Results show that the equilibrium melting temperatures (determined via either Hoffman¡VWeeks or Gibbs¡VThomson plots) of the copolymers are significantly lower than that of the corresponding sPS homopolymer. The PLM observations indicate that the axialitic growth rates in copolymers are drastically lower than that of the corresponding homopolymer at comparable backbone length and supercooling. Both XRD and TEM results indicate preferred formation of the ?" phase upon melt crystallization in the bulk state; however, the ?" phase (instead of ?' phase that is the more commonly observed for sPS homopolymers in the bulk state) is dominant in thin films.

Thesis (Ph. D.)--Auburn University, 2008. / Abstract. Vita. The following patent resulted from the dissertation research: Davis, V., Simonian, A.L., Nepal, D., Balasubramanian, S, "Preparation of Precisely Controlled Thin Film Nanocomposites of Carbon Nanotubes and Biomaterials", U.S. Provisional Patent Application No. 61/000,938, filed on 30 October 2007. The following peer-reviewed publications resulted from the dissertation research: Dhriti Nepal, Shankar Balasubramanian, Aleksandr Simonian, and Virginia Davis, "Mechanically Strong Antibacterial Thin Film Based on Single-Walled Carbon Nanotubes Armored with Biopolymers", Nano Letters ASAP article, May 2008 (# equal contribution) -- Shankar Balasubramanian, Iryna B. Sorokulova, Vitaly J. Vodyanoy, and Aleksandr L. Simonian, "Lytic Phage as a Specific and Selective Probe For Detection of Staphylococcus Aureus: A Surface Plasmon Resonance Spectroscopic Study", Biosensors and Bioelectronics, 2007, 22, 948-955 -- Shankar Balasubramanian, Alexander Revzin, Aleksandr Simonian, "Electrochemical Desorption of Proteins from Gold Electrode Surface", Electroanalysis, 2006, 18, 1885-1892 (Invited article) -- Vishwaprakash Nanduri, Shankar Balasubramanian, Srinivas Sista, Vitaly J. Vodyanoy, and Aleksandr L. Simonian, "Highly Sensitive Phage-based Biosensor for the Detection of ß-galactosidase", Analytica Chimica Acta, 2007, 589, 166- 172 -- H. Luckarift, Shankar Balasubramanian, S. Paliwal, G. Johnson and A. Simonian, "Enzyme-Encapsulated Silica Monolayers For Rapid Functionalization of a Gold Surface", Colloids and Surfaces B: Biointerfaces, 2007, 58, 28-33 (Invited article) -- Dong Wei, Omar Oyarzabal, Tung-Shi Huang, Shankar Balasubramanian, Srinivas Sista, Aleksandr Simonian, "Development of Surface Plasmon Resonance Biosensor For The Identification of Campylobacter jejuni", Journal of Microbiological Methods, 2007, 69, 78-85. The following conferences presentations resulted from the dissertation research: Covalent Immobilization of Organophosphorus Hydrolase on Carbon Nanotubes for Biosensor Applications, accepted for oral presentation at 12th International Meeting on Chemical Sensors, Jul. 13-16, 2008, Columbus, OH -- Electrochemical characteristics of SWNT-biopolymer nanocomposites, accepted for 213th meeting of The Electrochemical Society, May 18-23, 2008, Phoenix, AR -- Mechanically Robust Antibacterial Thin Films Composed of Single-Walled Carbon Nanotubes and Biopolymers, 2008 AIChE Spring National Meeting, Apr. 6-10, New Orleans, LA -- Production and characterization of protein and DNA based single wall carbon nanocomposites by layer-by-layer assembly, MRS Fall Meeting, Nov. 26-30, 2007, Boston, MA -- Gold surface modified with enzyme-encapsulated silica monolayers for biosensor application, The 58th Southeast Regional Meeting of the American Chemical Society, Nov. 1-4, 2006, Augusta, GA -- Electrochemical modulation of biological interfaces, 209th meeting of The Electrochemical Society, May 7-12, 2006, Denver, CO -- SPR based biosensor using lytic phage as a specific and selective probe for staphylococcus aureus detection, 57th Pittsburgh Conference on Analytical Chemistry and Applied Spectroscopy, Mar. 12-17, 2006, Orlando, FL -- Specific & selective detection of staphylococcus aureus by lytic phage using SPR biosensor, 57th Southeast / 61st Southwest Joint Regional Meeting of the American Chemical Society, Nov. 1-4, 2005, Memphis, TN -- Prevention of non-specific binding as a way to increase sensitivity of SPR-based sensors, 206th meeting of The Electrochemical Society, October 3-8, 2004, Honolulu, HI. Includes bibliographical references.

Made available in DSpace on 2014-06-11T19:26:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-09Bitstream added on 2014-06-13T19:13:48Z : No. of bitstreams: 1 silva_es_me_botfca.pdf: 2204058 bytes, checksum: 3b9b852e779ccf23c221c712179cfa66 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O controle químico tem sido utilizado freqüentemente com baixa eficiência para o fungo Didymella bryoniae, causador da doença crestamento gomoso ou cancro da haste em meloeiro. Porém, alternativas de controle como uso da enxertia e manipulação da nutrição devem ser estudadas. Avaliou-se porta-enxertos e concentrações de potássio na resistência de meloeiro rendilhado a Didymella bryoniae em ambiente protegido. No primeiro experimento, quatro porta-enxertos (melão „Dinero‟, melancia „Ojakkyo‟, abóbora „Strong Tosa‟ e cabaça „Calabash TI-191‟) foram inoculados com discos de micélio (7 mm de diâmetro) de quatro isolados de Didymella bryoniae (Dbr 34, Dbr 36, Dbr 37 e Dbr 39) em caule previamente ferido. Baseado no tamanho da lesão, o porta-enxerto „Dinero‟ foi susceptível, „Strong Tosa‟ moderadamente resistente, „Ojakkyo‟ e „Calabash TI-191‟ resistentes e o isolado Dbr 37 se mostrou mais agressivo dentre os isolados. No segundo experimento, o híbrido de meloeiro „Bônus II‟ foi enxertado nos porta-enxertos „Dinero‟ e „Strong Tosa‟, inoculadas com o isolado Dbr 37 do fungo e submetidas a diferentes concentrações de potássio (0; 62,5; 125; 187,5; 250 mg L-1), tanto plantas enxertadas como pé-franco. As plantas do híbrido „Bônus II‟ enxertadas foram resistentes ao fungo e as pé-franco susceptíveis. As concentrações de potássio não influenciaram no tamanho da lesão e na sobrevivência das plantas, porém influenciaram na condutância estomática e transpiração das plantas. / Chemical control of the fungus Didymella bryoniae, causal agent of gummy stem blight of melon, is frequently inefficient. Alternative methods as grafting on resistant rootstocks and nutrition handling must be studied. Different rootstocks and potassium concentrations were tested to control the disease on net melon grown under plastic house conditions. In a first experiment four rootstocks (melon „Dinero‟, watermelon „Ojakkyo‟, squash „Strong Tosa‟ and calabash „TI-191‟) were tested for resistance inoculated by disks mycelium (7 mm diameter) with four isolates ( Dbr 34, Dbr 36, Dbr 37 and Dbr 39) of D. bryoniae in stem previously injured. Based on lesion size „Dinero‟ proved to be susceptible to the fungus, „Strong Tosa‟ moderately resistant and „Ojakkyo‟ and „TI-191‟ resistant and the isolated Dbr 37 proved more aggressive among isolates. In a second experiment hybrid net melon „Bonus II‟ was grafted on rootstocks „Dinero‟ and „Strong Tosa‟, inoculated with isolate Dbr 37 of the fungus and submitted to different potassium concentration (0; 62,5; 125; 187,5; 250 mg L-1), inoculated both grafted plants as non grafted. Grafted plants of „Bonus II‟ were resistant to the fungus and the non grafted ones susceptible. Potassium concentrations did not influenced size of the lesion on the stem and also in the plants survival, however influence on stomatal conductance and transpiration plants.

Binary mixtures of polymers and soft nanocolloids, also called as polymer nanocomposites are well known and studied for their enormous potentials on various technological fronts. In this thesis blends of polystyrene grafted gold nanoparticles (PGNPs) and polystyrene (PS) are studied experimentally, both in bulk and in thin films. This thesis comprises three parts; 1) evolution of microscopic dynamics in the bulk(chapter-3),2) dispersion behavior of PGNPs in thin and ultra thin polymer matrices (chapter-4) 3) effect of dispersion on the glass transition behavior (chapter-5). In first part, the state of art technique, x-ray photon correlation spectroscopy is used to study the temperature and wave vector dependent microscopic dy¬namics of PGNPs and PGNP-PS mixtures. Structural similarities between PGNPs and star polymers (SPs) are shown using small angle x-ray scatter¬ing and scaling relations. We find unexpected (when compared with SPs) non-monotonic dependence of the structural relaxation time of the nanoparticles with functionality (number of arms attached to the surface). Role of core-core attractions in PGNPs is shown and discussed to be the cause of anomalous behavior in dynamics. In PGNP-PS mixtures, we find evidence of melting of the dynamically arrested state of the PGNPs with addition of PS followed by a reentrant slowing down of the dynamics with further increase in polymer frac¬tion, depending on the size ratio(δ)of PS and PGNPs. For higher δ the reen¬trant behavior is not observed with polymer densities explored here. Possible explanation of the observed dynamics in terms of the presence of double-glass phase is provided. The correlation between structure and reentrant vitrifica¬tion in both pristine PGNPs and blends are derived rather qualitatively. In the second part, the focus is shifted to miscibility between PGNPs and polymers under confinement i.e., in thin films. This chapter provide a compre¬hensive study on the different parameters affecting dispersion viz., annealing conditions, fraction of the added particles, polymer-particle interface and more importantly the thickness of the films. Changes in the dispersion behavior with annealing is shown and the need for annealing the films at temperatures higher than the glass transition temperature of the matrix polymers is clearly elucidated. Irrespective of the thickness of the films( 20 and 65 nm) studied, immiscible particle-polymer blends unequivocally prove the presence of gradi¬ent in dynamics along the depth of the films. To our knowledge for the first time, we report results on confinement induced enhancement in the dispersion of the nanoparticles in thin polymer films. The enhanced dispersion is argued to be facilitated by the increased free volume in the polymer due to confinement as shown by others. Based on these results we have proposed a phase diagram for dispersibility of the nanoparticles in polymer films. The phase diagram for ultra thin films highlights an important point: In ultra thin films the particles are dispersed even with grafting molecular weight less than matrix molecular weight. In the third part, we have studied the glass transition of the thin films whose structure has been studied earlier in the earlier part. Non-monotonic variation in glass transition with the fraction of particles in thin films has increased our belief on the gradient in the dynamics of thin polymer films. En¬hanced dispersion with confinement is captured with the enhanced deviation in glass transition temperature of ultra thin films. Effect of miscibility param¬eter on Tgis studied and the results are explained with the subtle interplay of polymer-particle interface and confinement.